Practical limits to column performance in liquid chromatography - Optimal operations.

Abstract

Columns of different structures have different potential kinetic performance - the trade-off between separation, time, and pressure. However, the full potential of a structure cannot always be realized in practically existing columns. Each combination of column efficiency, time, and pressure requires certain cross-sectional dimensions of the column flow-through channels. However, there are limits to the narrowest flow-through channels that can be manufactured with current technology. As a result, columns of some structures cannot be optimized for providing the required efficiency in the shortest time. Additionally, the full potential of its structure can be realized only if a column can operate at the highest pressure available from liquid chromatography (LC) equipment, has sufficient loadability, and satisfies other practical requirements. Equations tailored for a systematic approach to evaluation of factors affecting performance of optimized LC columns (effects of column structure, column dimensions, operational conditions, etc.) were developed. Parameters quantifying the performance of a specific column at and below its largest acceptable pressure were identified. New objective performance parameters of columns and their structures were introduced. Among them are the apparent structural quality factor accounting for the effect of insufficiently high pressure acceptable for the column, the dimensionless plate duration - the parameter of a column structure affecting its performance when the pressure is not limited, - and others. Applying the theory developed herein to published data, the performance of several differently structured columns is evaluated, and the factors affecting their comparative performance are discussed. In the final count, not the quality of a column structure, but practical factors such as the narrowest manufacturable flow-through channels can dominate the choice of the kinetically most suitable column for a practical LC analysis.